A new method for identifying proteins involved in DNA methylation through reverse genetics in Arabidopsis

Copyright © 2024. Published by Elsevier B.V.

Bibliographische Detailangaben
Veröffentlicht in:Plant science : an international journal of experimental plant biology. - 1985. - 352(2025) vom: 16. März, Seite 112376
1. Verfasser: Miao, Wei (VerfasserIn)
Weitere Verfasser: Dai, Jie, Zhang, Li, Liang, Zhile, Sun, Xiaoxuan, Huang, Meizi, Zhang, Aqin, Zheng, Long, Li, Yongjun, Li, Ying
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2025
Zugriff auf das übergeordnete Werk:Plant science : an international journal of experimental plant biology
Schlagworte:Journal Article CRISPR DNA methylation Gene co-expression Reduced luminescence 1 (rl1) Reverse genetics Arabidopsis Proteins ROS1 protein, Arabidopsis Nuclear Proteins
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520 |a Forward genetic screens have uncovered numerous genes involved in DNA methylation regulation, but these methods are often time-intensive, costly, and labor-intensive. To address these limitations, this study utilized CRISPR technology to knockout selected co-expressed genes, enabling the rapid identification of low luciferase (LUC) luminescence mutants in the Col-LUC line, which harbors a LUC transgene driven by a 2 × 35S promoter in Arabidopsis. As proof of concept, the repressor of silencing 1 (ROS1) and RNA-directed DNA methylation 1 (RDM1) genes were used as controls, while the increased DNA methylation 3 (IDM3) gene, co-expressed with ROS1, was selected as the target for gene knockout experiments. The results demonstrated that combining co-expression analysis with CRISPR technology is an effective strategy for generating low LUC luminescence mutants in the Col-LUC line. Notably, a new mutant, named reduced luminescence 1 (rl1), was identified through this approach. The rl1 mutant exhibited genome-wide DNA hypermethylation, and its reduced luminescence phenotype was largely reversed by treatment with the DNA methylation inhibitor 5-Aza-2'-deoxycytidine, confirming its anti-silencing role in DNA methylation regulation. This study presents a novel and efficient approach for obtaining low luminescence mutants in the Col-LUC line and identifies RL1 as a previously uncharacterized protein involved in DNA methylation regulation 
650 4 |a Journal Article 
650 4 |a CRISPR 
650 4 |a DNA methylation 
650 4 |a Gene co-expression 
650 4 |a Reduced luminescence 1 (rl1) 
650 4 |a Reverse genetics 
650 7 |a Arabidopsis Proteins  |2 NLM 
650 7 |a ROS1 protein, Arabidopsis  |2 NLM 
650 7 |a Nuclear Proteins  |2 NLM 
700 1 |a Dai, Jie  |e verfasserin  |4 aut 
700 1 |a Zhang, Li  |e verfasserin  |4 aut 
700 1 |a Liang, Zhile  |e verfasserin  |4 aut 
700 1 |a Sun, Xiaoxuan  |e verfasserin  |4 aut 
700 1 |a Huang, Meizi  |e verfasserin  |4 aut 
700 1 |a Zhang, Aqin  |e verfasserin  |4 aut 
700 1 |a Zheng, Long  |e verfasserin  |4 aut 
700 1 |a Li, Yongjun  |e verfasserin  |4 aut 
700 1 |a Li, Ying  |e verfasserin  |4 aut 
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